Department of Ophthalmology, School of Medicine, Trakya University - Balkan Campus, Edirne, Turkey.
Department of Medical Biology, School of Medicine, Trakya University - Balkan Campus, Edirne, Turkey.
Cutan Ocul Toxicol. 2021 Jun;40(2):103-114. doi: 10.1080/15569527.2021.1902343. Epub 2021 Apr 8.
The main characteristic of proliferative vitreoretinopathy (PVR) is migration, adhesion, and epithelial-mesenchymal transition (EMT) of retinal pigment epithelial cells (RPE). Eupatilin is a naturally occurring flavone that has the potential to inhibit cell proliferation and EMT. However, its efficacy on the PVR model induced by transforming growth factor-2 (TGF-β2) is unknown. In this study, the potential effect of eupatilin on proliferation and EMT in the treatment of RPE was investigated.
Serum starved human RPE cells (ARPE-19) were treated with 10 ng/ml TGF-β2 alone or co-treated with 25 μM eupatilin for 48 h. Quantitative real-time PCR and Western blot analysis were used to assess targets at the mRNA and protein expression level, respectively. Apoptosis and cell cycle progression was assessed by image-based cytometry. The effect of treatment on cell migration was evaluated by wound healing assay.
Eupatilin inhibited TGF-β2-induced RPE cell proliferation via regulating the cell cycle and inducing apoptosis. TGF-β2 upregulated mRNA expression of mesenchymal markers fibronectin and vimentin was significantly downregulated by the treatment, while the epithelial markers E-cadherin and occludin expression was upregulated. The therapy significantly suppressed TGF-β2 encouraged cell migration through downregulating the expression of transcription factors Twist, Snail, and ZEB1 induced by TGF-β2. Furthermore, eupatilin significantly inhibited the expression of MMP-1, -7, and -9, and suppressed NF-κB signalling.
These results suggest that eupatilin could inhibit the proliferation and transformation into fibroblast-like cells of RPE cells; thus the agent may be a potential therapeutic value in treating PVR.
增生性玻璃体视网膜病变（PVR）的主要特征是视网膜色素上皮细胞（RPE）的迁移、黏附和上皮-间充质转化（EMT）。芹菜素是一种天然存在的类黄酮，具有抑制细胞增殖和 EMT 的潜力。然而，其在转化生长因子-2（TGF-β2）诱导的 PVR 模型中的疗效尚不清楚。在这项研究中，研究了芹菜素在治疗 RPE 中对增殖和 EMT 的潜在影响。
用 10ng/ml TGF-β2 单独或与 25μM 芹菜素共同处理血清饥饿的人 RPE 细胞（ARPE-19）48h。分别采用定量实时 PCR 和 Western blot 分析评估 mRNA 和蛋白表达水平的靶标。通过基于图像的细胞计数评估细胞凋亡和细胞周期进程。通过划痕愈合试验评估治疗对细胞迁移的影响。
芹菜素通过调节细胞周期和诱导细胞凋亡抑制 TGF-β2 诱导的 RPE 细胞增殖。TGF-β2 上调了间充质标志物纤维连接蛋白和波形蛋白的 mRNA 表达，经治疗后显著下调，而上皮标志物 E-钙黏蛋白和封闭蛋白的表达上调。该疗法通过下调 TGF-β2 诱导的转录因子 Twist、Snail 和 ZEB1 的表达，显著抑制了 TGF-β2 促进的细胞迁移。此外，芹菜素还显著抑制了 MMP-1、-7 和 -9 的表达，并抑制了 NF-κB 信号通路。
这些结果表明，芹菜素可以抑制 RPE 细胞的增殖和向成纤维样细胞的转化；因此，该药物可能在治疗 PVR 方面具有潜在的治疗价值。
